Basic NeuroscienceSelf-built microdialysis probes with improved recoveries of ATP and neuropeptides

- The design of a custom-made microdialysis probe is presented step by step.
- Three different probes were compared for recoveries.
- As analytes, we chose glucose, acetylcholine, ATP and three neuropeptides.
- Recovery of the custom-made 30 kDa probe is highest and most stable.
- The use of ZipTip® μC-18 collection tips improves the purity of the sample.

BackgroundMicrodialysis is an established technique for collecting small molecular weight substances (e.g. neurotransmitter and energy metabolites) from the extracellular space. The major element of microdialysis is the probe which contains a semi-permeable membrane and is exposed to the interstitial space. As the microdialysis technique has major advantages, e.g. versatility and use in awake animals, commercially produced probes are in great demand.New methodWe here present the design of a probe assembly step by step which will enable researchers to build custom-made probes. Probe recoveries of substances with different molecular weight (ranging from 100 to 1600 Da) were compared for three different probes (CMA 12 Elite probe, custom-made 10 kDa and 30 kDa probes). Recoveries of glucose, lactate, acetylcholine, choline, ATP and the neuropeptides angiotensin II, substance P and somatostatin are presented.ResultsWe found that the 10 kDa probe is only useful for compounds up to 1000 Da while recoveries of the CMA-12 Elite Probe are variable and apparently dependent on ionic charges of analytes. The recovery of the custom-made 30 kDa probe is highest and evidently not influenced by physicochemical parameters of analytes. In a further optimization step, we describe the use of ZipTip® μC-18 collection tips to replace the outlet tubing when purifying the dialysate for MALDI-MS measurements of neuropeptides.Comparison with existing methodsThe results show that self-built microdialysis probes can be equally or more effective than commercially available probes.ConclusionsSelf-built microdialysis probes with large pore-membranes are capable of dialyzing ATP and neuropeptides.